{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,4]],"date-time":"2026-04-04T11:37:28Z","timestamp":1775302648240,"version":"3.50.1"},"reference-count":9,"publisher":"S. Karger AG","issue":"2","license":[{"start":{"date-parts":[[2004,1,1]],"date-time":"2004-01-01T00:00:00Z","timestamp":1072915200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/www.karger.com\/Services\/SiteLicenses"},{"start":{"date-parts":[[2004,1,1]],"date-time":"2004-01-01T00:00:00Z","timestamp":1072915200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/www.karger.com\/Services\/SiteLicenses"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Fetal Diagn Ther"],"published-print":{"date-parts":[[2004]]},"abstract":"<i>Objective:<\/i> Spinal muscular atrophy (SMA) is a common autosomal recessive neuromuscular disorder caused in most patients by homozygous deletion of the <i>SMN1 <\/i>gene. For a carrier couple at a 25% risk of affected offspring, preimplantation genetic diagnosis (PGD) offers an alternative to prenatal diagnosis and termination of affected pregnancies. Our objective was to develop an accurate and reliable single-cell multiplex nested PCR analysis for PGD of SMA. <i>Methods:<\/i> The method was developed on single blood leukocytes, obtained from healthy controls and an adult SMA type III patient with a known homozygous deletion of <i>SMN1 <\/i>exon 7 and 8. Multiplex nested PCR on single cells was used to co-amplify exons 7 and 8 of <i>SMN<\/i>. Additional multiplexing was performed with the <i>ZFX\/ZFY <\/i>gene for sexing. Following successful establishment of the multiplex nested PCR protocol in single leukocytes, the technique was employed for PGD in 4 patients for a total of 7 cycles. In 2 patients, sexing was simultaneously performed using <i>ZFX\/ZFY<\/i>. <i>Results:<\/i> 220 single leukocytes from a normal individual and 220 from an SMA patient were analyzed. Exon 7 of <i>SMN1<\/i> was amplified in 99% of normal single leukocytes and in none of the SMA-affected leukocytes. Exon 7 of <i>SMN2<\/i> was amplified in 100% of both normal and SMA-affected leukocytes. Exon 8 of <i>SMN1<\/i> was amplified in 98% of normal cells and in none of the SMA-affected leukocytes. Exon 8 of <i>SMN2<\/i> was amplified in 96% of both normal and SMA-affected leukocytes. Amplification efficiency was 99% for <i>ZFX\/ZFY<\/i>. There were no false-negative results and no contamination was detected in all wash-drop blanks tested. Seven PGD cycles were performed in 4 SMA-carrier couples with successful molecular analysis of 34 embryos and a total of 15 normal embryos transferred in 7 cycles. One clinical pregnancy has resulted in the delivery of a healthy male. Amniocentesis performed at 17 weeks confirmed the correct diagnosis for both SMA and sexing. <i>Conclusions:<\/i> These results suggest that our multiplex nested PCR protocol offers an efficient and accurate method for PGD of SMA while enabling the simultaneous analysis of an additional loci.<\/jats:p>","DOI":"10.1159\/000075151","type":"journal-article","created":{"date-parts":[[2004,2,6]],"date-time":"2004-02-06T10:57:01Z","timestamp":1076065021000},"page":"199-206","source":"Crossref","is-referenced-by-count":20,"title":["Multiplex Nested PCR for Preimplantation Genetic Diagnosis of Spinal Muscular Atrophy"],"prefix":"10.1159","volume":"19","author":[{"given":"Mira","family":"Malcov","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tamar","family":"Schwartz","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nava","family":"Mei-Raz","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Dalit Ben","family":"Yosef","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ami","family":"Amit","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Joseph B.","family":"Lessing","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ruth","family":"Shomrat","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Avi","family":"Orr-Urtreger","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yuval","family":"Yaron","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"127","published-online":{"date-parts":[[2004,2,13]]},"reference":[{"key":"ref1","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(80)90847-8"},{"key":"ref2","doi-asserted-by":"publisher","DOI":"10.1038\/344540a0"},{"key":"ref3","doi-asserted-by":"publisher","DOI":"10.1016\/0140-6736(90)91803-I"},{"key":"ref4","doi-asserted-by":"publisher","DOI":"10.1006\/geno.1996.0147"},{"key":"ref5","doi-asserted-by":"publisher","DOI":"10.1016\/0092-8674(95)90061-6"},{"key":"ref6","doi-asserted-by":"publisher","DOI":"10.1086\/302369"},{"key":"ref7","doi-asserted-by":"publisher","DOI":"10.1016\/S0168-9525(97)01166-9"},{"key":"ref8","doi-asserted-by":"publisher","DOI":"10.1002\/pd.110"},{"key":"ref9","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(95)90732-7"}],"container-title":["Fetal Diagnosis and Therapy"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.karger.com\/Article\/Pdf\/75151","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.karger.com\/Article\/Pdf\/75151","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,4,24]],"date-time":"2025-04-24T03:59:38Z","timestamp":1745467178000},"score":1,"resource":{"primary":{"URL":"https:\/\/karger.com\/article\/doi\/10.1159\/000075151"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2004]]},"references-count":9,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2004,4,1]]}},"URL":"https:\/\/doi.org\/10.1159\/000075151","archive":["Portico"],"relation":{},"ISSN":["1015-3837","1421-9964"],"issn-type":[{"value":"1015-3837","type":"print"},{"value":"1421-9964","type":"electronic"}],"subject":[],"published":{"date-parts":[[2004]]}}}